| 质粒名称: | pET15b_Bst-LF |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 153313 |
| 存储实验室: | Drew Endy, Philippa Marrack |
|
载体骨架基本信息 |
|
| 载体名称: | pET-15b |
| 完整质粒大小: | 7473bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Bst Large Fragment |
| 物种来源: | 其他 |
|
文献引用方法 |
|
| 材料和方法部分: | pET15b_Bst-LF was a gift from Drew Endy & Philippa Marrack (Addgene plasmid # 153313 ; http://n2t.net/addgene:153313 ; RRID:Addgene_153313) |
买家导航
pET15b_Bst-LF载体质粒基本信息
pET15b_Bst-LF质粒图谱载体图谱和pET15b_Bst-LF载体序列质粒序列多克隆位点信息
pET15b_Bst-LF质粒载体简介
pET15b_Bst-LF质粒序列载体序列
LOCUS pET15b_Bst-LF 7473 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression of the Bst Large Fragment.
ACCESSION .
VERSION .
KEYWORDS pET15b_Bst-LF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7473)
TITLE COVID-19 Diagnostic Toolkit Enzymes
REFERENCE 2 (bases 1 to 7473)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7473
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 14..33
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 117..136
/label=T7
/note="T7 promoter, forward primer"
promoter 117..135
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 136..160
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
RBS 175..197
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 216..233
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 243..260
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
primer_bind complement(2090..2108)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2104..2151
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(2326..2354)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(2381..2399)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2467..2571
/gene="bla"
/label=AmpR promoter
CDS 2572..3432
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(2790..2809)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3603..4191
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4092..4111
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4345..4362
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4377..4516
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4442..4461)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4602..4624
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4618..4809)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6121..7203)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 7165..7184
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7204..7281)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
61 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
121 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
181 taactttaag aaggagatat accatgggca gcagccatca tcatcatcat cacagcagcg
241 gcctggtgcc gcgcggcagc catagccata tgaccgacga aggcgaaaaa ccgctggcgg
301 gtatggactt tgcgatcgct gatagcgtta ccgacgagat gctggcagat aaagcagcgc
361 tagtagttga agttgtgggc gataactacc atcatgcgcc gatcgtgggt atcgctttgg
421 cgaacgagcg cggccgtttt ttcctgcgcc cggaaaccgc actggcggac ccgaagtttc
481 tggcttggct gggcgacgaa acgaaaaaaa agactatgtt cgattctaaa cgtgcagcgg
541 tggctctcaa atggaaaggt atcgaattgc gcggtgttgt gttcgacctg ctgctggctg
601 cgtacctgct cgatcctgct caggctgcgg gtgacgtggc agctgtagcg aaaatgcacc
661 agtacgaagc cgtacgctct gatgaagcgg tttatggtaa aggtgcgaaa cgtaccgtgc
721 cagacgaacc aaccctggcg gaacacctgg ctcgcaaagc agccgctatc tgggccctgg
781 aggagcctct gatggatgaa cttcgccgca acgaacagga ccgtctgctc acagaactgg
841 aacagccgtt ggcgggtatt ctggcaaaca tggagttcac tggtgtaaaa gtggacacta
901 agcgcctgga acaaatgggt gccgaactga ctgagcagct tcaggcagtt gaacgccgca
961 tctacgaact ggcgggtcag gagtttaaca tcaactctcc taaacagctg ggcaccgtgt
1021 tgttcgacaa actgcaactg ccggtgttga agaaaacgaa aaccggctac agcacctccg
1081 ccgatgttct ggaaaaactg gcaccgcatc acgaaatcgt tgaacacatc ctgcactacc
1141 gtcagctggg taaactgcaa tctacctaca tcgagggctt actgaaagta gtgcacccgg
1201 tgaccggtaa agtgcacact atgttcaacc aggctctgac gcagaccggc cgcctgtcct
1261 ccgttgaacc gaacctgcaa aacatcccca tccgtctgga agagggccgt aagatccgtc
1321 aggcattcgt gccgtctgaa ccggactggc tgattttcgc tgctgattac tctcaaatcg
1381 aactgcgtgt tcttgctcac attgcggagg acgataacct gattgaagca ttccgtcgtg
1441 gtctggacat tcacactaaa acggctatgg acattttcca tgtttccgaa gaagatgtga
1501 ccgctaatat gcgtcgtcag gctaaagcgg taaactttgg tatcgtatac ggtatctccg
1561 attacggcct ggcacagaac ctgaacatca cccgcaaaga agcggctgag tttatcgaac
1621 gttatttcgc ctcgttcccc ggtgttaaac agtacatgga taacatcgtt caagaagcta
1681 aacagaaggg ttatgtgacc accctgctgc accgtcgtcg ttacctgccg gacatcacct
1741 cccgtaactt caacgtgcgt tctttcgccg aacgtacggc tatgaatacc cctatccagg
1801 gcagcgcggc ggacatcatc aaaaaagcca tgatcgacct gagcgtgcgc ctgcgcgaag
1861 aacgtttgca ggctcgcctg cttctgcaag tgcacgatga gctgatccta gaagctccaa
1921 aagaggaaat tgaacgtttg tgccgtctgg ttccggaagt tatggaacag gctgtcgcac
1981 tgcgtgttcc gcttaaagtt gattaccatt acggtcccac ttggtacgac gccaagtgag
2041 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
2101 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
2161 ggaactatat ccggatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
2221 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
2281 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
2341 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
2401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
2461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
2521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
2581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
2641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
2701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
2761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
2821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
2881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
2941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
3001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
3061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
3121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
3181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
3421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
3481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
3541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
3601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
3661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
3721 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
3781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
3841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
3901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
3961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
4021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
4081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
4141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
4201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
4261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
4321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
4381 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
4441 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
4501 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
4561 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
4621 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
4681 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
4741 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
4801 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
4861 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
4921 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
4981 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
5041 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
5101 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
5161 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
5221 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
5281 ggacccaacg ctgcccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga
5341 tatgttctgc caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc
5401 aattcttgga gtggtgaatc cgttagcgag gtgccgccgg cttccattca ggtcgaggtg
5461 gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga caaggtatag ggcggcgcct
5521 acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc
5581 agcggtccaa tgatcgaagt taggctggta agagccgcga gcgatccttg aagctgtccc
5641 tgatggtcgt catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg
5701 ccggaagcga gaagaatcat aatggggaag gccatccagc ctcgcgtcgc gaacgccagc
5761 aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
5821 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
5881 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
5941 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
6001 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
6061 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
6121 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
6181 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
6241 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
6301 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
6361 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
6421 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
6481 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
6541 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
6601 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
6661 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
6721 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
6781 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
6841 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
6901 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
6961 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
7021 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
7081 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
7141 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
7201 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
7261 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
7321 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
7381 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
7441 aacaagcgct catgagcccg aagtggcgag ccc
//
